Abstract:
:Accurate and rapid identification of methicillin-resistant Staphylococcus aureus (MRSA) is needed to screen MRSA carriers and improve treatment. The current widely used duplex PCR methods are not able to differentiate MRSA from coexisting methicillin-susceptible S. aureus (MSSA) or other methicillin-resistant staphylococci. In this study, we aimed to develop a direct method for accurate and rapid detection of MRSA in clinical samples from open environments, such as nasal swabs. The new molecular assay is based on detecting the cooccurrence of nuc and mecA markers in a single bacterial cell by utilizing droplet digital PCR (ddPCR) with the chimeric lysin ClyH for cell lysis. The method consists of (i) dispersion of an intact single bacterium into nanoliter droplets, (ii) temperature-controlled release of genomic DNA (gDNA) by ClyH at 37°C, and (iii) amplification and detection of the markers (nuc and mecA) using standard TaqMan chemistries with ddPCR. Results were analyzed based on MRSA index ratios used for indicating the presence of the duplex-positive markers in droplets. The method was able to achieve an absolute limit of detection (LOD) of 2,900 CFU/ml for MRSA in nasal swabs spiked with excess amounts of Escherichia coli, MSSA, and other mecA-positive bacteria within 4 h. Initial testing of 104 nasal swabs showed that the method had 100% agreement with the standard culture method, while the normal duplex qPCR method had only about 87.5% agreement. The single-bacterium duplex ddPCR assay is rapid and powerful for more accurate detection of MRSA directly from clinical specimens.
journal_name
J Clin Microbioljournal_title
Journal of clinical microbiologyauthors
Luo J,Li J,Yang H,Yu J,Wei Hdoi
10.1128/JCM.00716-17subject
Has Abstractpub_date
2017-10-01 00:00:00pages
2946-2955issue
10eissn
0095-1137issn
1098-660Xpii
JCM.00716-17journal_volume
55pub_type
杂志文章abstract::A biphasic blood culture bottle (BiPB: GIBCO Laboratories, North Andover, Mass.) with an architectural design that physically separates the agar slant from the broth was compared with a conventional vented monophasic bottle (MPB-A) for use in the routine culture of blood. Both bottles contained tryptic soy broth. Tryp...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.23.4.700-703.1986
更新日期:1986-04-01 00:00:00
abstract::Amplified fragment length polymorphism (AFLP) was employed as a genetic analysis tool for the study of the genetic relatedness of Mycobacterium avium subsp. paratuberculosis isolates harvested from bovine fecal samples and from bovine or human tissues. This analysis revealed genetic differences between these two isola...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01123-10
更新日期:2011-06-01 00:00:00
abstract::Human and porcine enterotoxigenic strains of Escherichia coli were cultivated in tryptone-yeast extract medium or brain heart infusion broth and tested for production of heat-labile enterotoxin by the GM1 ganglioside enzyme-linked immunosorbent assay (GM1-ELISA) and the Y1 adrenal cell assay. When testing for enteroto...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.15.2.298-301.1982
更新日期:1982-02-01 00:00:00
abstract::We conducted a case-control study using the Fungitell assay, the novel Platelia Candida Antigen (Ag) Plus and Candida Antibody (Ab) Plus assays, and the Cand-Tec latex agglutination test to evaluate the usefulness of (1→3)-β-D-glucan (BDG), mannan antigen with/without anti-mannan antibody, and Cand-Tec Candida antigen...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.02473-12
更新日期:2013-04-01 00:00:00
abstract::The epidemiological implications of the recent separation of "Entamoeba histolytica" into two separate species, pathogenic E. histolytica sensu stricto and commensal E. dispar, will not become apparent without methods of distinguishing between them which are applicable to large numbers of specimens. We have modified a...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.35.5.1108-1111.1997
更新日期:1997-05-01 00:00:00
abstract::We report a fatal case of acute lower respiratory tract disease with human rhinovirus C (HRV-C) as the unique cause in a 19-month-old girl with a history of repeated episodes of bronchiolitis. HRV-C type 8 nucleic acids were observed in respiratory, stool, and cerebrospinal fluid samples, and infectious virions were i...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.03484-14
更新日期:2015-05-01 00:00:00
abstract::A premarket trial of the Becton Dickinson Directigen respiratory syncytial virus membrane-based enzyme immunoassay compared the test with virus isolation for the detection of respiratory syncytial virus in 583 nasopharyngeal aspirates. After modification, the Directigen test showed a sensitivity of 83% and a specifici...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.28.6.1458-1459.1990
更新日期:1990-06-01 00:00:00
abstract::The specificities and sensitivities of the Bactec mycobacterial growth indicator tube (MGIT) system for the recovery of Mycobacterium tuberculosis from pleural fluid are not statistically different than those of the Myco/F lytic liquid culture system. The time to positivity is shorter in the MGIT system (12.7 versus 2...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.02742-14
更新日期:2015-04-01 00:00:00
abstract::Streptococcus uberis and Streptococcus parauberis reference strains and isolates obtained from routine diagnostics were investigated by PCR with oligonucleotide primers designed according to species-specific parts of the 16S rRNA gene, the 23S rRNA gene, and the 16S-23S rRNA intergenic spacer region of both species. A...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.39.4.1618-1621.2001
更新日期:2001-04-01 00:00:00
abstract::Moxalactam is highly hydrolyzed by plasmid-mediated metallo-β-lactamases (MBLs), whereas it is poorly inactivated by serine-active carbapenemases. This study demonstrated that moxalactam resistance constituted an effective screen for MBL expression in enterobacteria, which could be confirmed, even in low-MBL-producing...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00328-11
更新日期:2011-07-01 00:00:00
abstract::An enzyme-linked immunosorbent assay was developed for the detection of circulating free and complexed staphylococcal antigens in various clinical categories of staphylococcal infections. Circulating immune complexes were studied by the polyethylene glycol precipitation method. Circulating immune complexes and staphyl...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.16.6.1019-1024.1982
更新日期:1982-12-01 00:00:00
abstract::A reverse transcription-PCR (RT-PCR) assay based on automated fluorescent capillary electrophoresis and GeneScan software analysis was developed to detect six common respiratory viruses in clinical specimens from young children. Assays for human respiratory syncytial virus (HRSV); human parainfluenza viruses 1, 2, and...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.41.9.4298-4303.2003
更新日期:2003-09-01 00:00:00
abstract::Malaria is one of the leading causes of infectious disease in travelers returning from the tropics. The diagnosis of malaria is typically performed by examining Giemsa-stained thick and thin peripheral blood smears, which is time consuming, labor intensive, and requires high levels of proficiency. Alternatively, loop-...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00806-17
更新日期:2017-10-01 00:00:00
abstract::We report an uncommon clinical presentation of a unique case of fatal invasive fungal cerebral vasculitis due to Arthrographis kalrae in a nonimmunocompromised host. The identity of the fungus was determined by morphological characteristics and by analysis of internal transcribed spacer 1 sequences and was confirmed b...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00800-08
更新日期:2008-09-01 00:00:00
abstract::Plasmodium nucleic acids have been detected in serum and plasma, but there is little published data describing the diagnostic performance of malaria nucleic acid amplification tests (NAATs) using these specimen types. Previously, our group described a multiplex NAAT for the detection of dengue virus, Leptospira, and P...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01876-15
更新日期:2015-11-01 00:00:00
abstract::The genus Corynebacterium is a heterogeneous group of species comprising human and animal pathogens and environmental bacteria. It is defined on the basis of several phenotypic characters and the results of DNA-DNA relatedness and, more recently, 16S rRNA gene sequencing. However, the 16S rRNA gene is not polymorphic ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.42.9.3925-3931.2004
更新日期:2004-09-01 00:00:00
abstract::A bacitracin (0.04 units, BBL)-SXT (trimethoprim, 1.25 mg, plus sulfamethoxazole, 23.75 mg, BBL) susceptibility test was 94% accurate in presumptively identifying streptococci as either group A, B or not group A and B. ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:
更新日期:1976-08-01 00:00:00
abstract::The objective of this study was to evaluate the clinical characteristics and outcomes of hospitalized patients tested for Clostridium difficile and determine the correlation between pretest probability for C. difficile infection (CDI) and assay results. Patients with testing ordered for C. difficile were enrolled and ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.02126-16
更新日期:2017-02-01 00:00:00
abstract::Data pertaining to 20 strains of Yensinia enterocolitica isolated from the cecal content of swine slaughtered at Toronto Abattoirs are presented. Fifteen of 20 cultures belonged to the two predominant human serotypes in Ontario (i.e., serotype O:3 and O:5,27.) Seven cultures of Y. enterocolitica O:3 belonged to the "C...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:
更新日期:1975-12-01 00:00:00
abstract::The increasing number of multidrug-resistant Mycobacterium tuberculosis strains has stimulated the interest of investigators in finding a rapid method for susceptibility testing. We used commercially available rRNA DNA-bioluminescence-labelled probes (Accu-Probe, Gen Probe, Inc. San Diego, Calif.) for this purpose. Th...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.35.10.2521-2525.1997
更新日期:1997-10-01 00:00:00
abstract::Colistin-nalidixic acid agar, although recently recommended as a replacement for blood agar for primary plating of urine specimens ( Fung et al., J. Clin. Microbiol. 16:632-636, 1982), has also been reported to suppress the growth of some strains of staphylococci that are susceptible to colistin (polymyxin E). The sus...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.19.5.714-716.1984
更新日期:1984-05-01 00:00:00
abstract::Cag pathogenicity island-containing Helicobacter pylori (type I) induces signal transduction pathways resulting in tyrosine phosphorylation of proteins adjacent to the site of bacterial adhesion on host gastric epithelial cells. Conventional block PCR-restriction fragment length polymorphism (RFLP) and real-time Light...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.41.7.3112-3118.2003
更新日期:2003-07-01 00:00:00
abstract::We evaluated the Prodesse ProFlu-1 real-time reverse transcription-PCR multiplex assay with the SmartCycler instrument for the detection of human respiratory syncytial virus (RSV) and influenza A and B viruses in comparison to conventional cell culture and antigen immunoassays with the BD Directigen A+B and Binax NOW ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01213-08
更新日期:2009-03-01 00:00:00
abstract::A novel PCR restriction analysis method using the RNA polymerase beta-subunit- coding gene (rpoB) was employed to both detect and identify Helicobacter pylori in biopsy specimens and culture isolates. The rpoB DNAs (458 bp) were specifically amplified by PCR with the Helicobacter-specific primers (HF and HR). Based on...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.41.7.3387-3391.2003
更新日期:2003-07-01 00:00:00
abstract::We report a case of cerebral phaeohyphomycosis in a 36-year-old male caused by the neurotropic fungus Ramichloridium obovoideum (Matushima) de Hoog 1977 (Ramichloridium mackenziei Campbell et Al-Hedaithy 1993). This man resided in the Middle East, where the fungus appears to be endemic and, possibly, geographically re...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章,评审
doi:10.1128/JCM.36.3.708-715.1998
更新日期:1998-03-01 00:00:00
abstract::In this study, 102 herpes simplex virus isolates were typed by cell culture selection (chicken embryo cells and guinea pig embryo cells [CE/GPE]), (E)-5-(2-bromovinyl)-2'-deoxyuridine (BVDU) sensitivity, plaque reduction neutralization, and indirect immunofluorescence staining techniques. The percentages of agreement ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.17.3.396-399.1983
更新日期:1983-03-01 00:00:00
abstract::Since the late 1980s several studies have described the increased incidence and severity of invasive group A streptococcal (GAS) infections. However, most studies on GAS pathogenesis have focused on information obtained during outbreaks. We analyzed isolate distribution and host susceptibility as part of a nationwide ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.43.7.3101-3109.2005
更新日期:2005-07-01 00:00:00
abstract::The cellular fatty acid compositions and ubiquinone contents of 182 Legionella strains representing 23 species were determined by capillary gas-liquid chromatography and reverse-phase high-performance liquid chromatography, respectively. Except for the type strain of Legionella erythra (ATCC 35303T), all Legionella sp...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.27.3.465-473.1989
更新日期:1989-03-01 00:00:00
abstract::A rapid method for detection of respiratory syncytial virus (RSV) in nasopharyngeal aspirates, involving a combination of reverse transcription and polymerase chain reaction amplification (RT-PCR), has been developed. The RT-PCR assay employs oligonucleotide primers specific for the region of the RSV genome which enco...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.30.4.901-904.1992
更新日期:1992-04-01 00:00:00
abstract::Sequential specimens from nine allograft recipients were examined by using a variety of methods to detect primary cytomegalovirus (CMV) infection as rapidly as possible posttransplantation. Sera were examined for immunoglobulin G (IgG) and IgM antibodies by immunoblotting, enzyme immunoassay, and immunofluorescence an...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.27.12.2672-2677.1989
更新日期:1989-12-01 00:00:00